Protein O-Fucosyltransferase 1 Undergoes Interdomain Flexibility in Solution
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GlycosyltransferasesO-fucosylationProtein dynamicsAtomic force microscopySinglemolecule methods
Lira-Navarrete, E.; Pallarés, M.C.; Castello, F.; Ruedas-Rama, M.J.; Orte, A.; Lostao, A.; Hurtado-Guerrero, R. Protein O-Fucosyltransferase 1 Undergoes Interdomain Flexibility in Solution. Molecules 2021, 26, 2105. https:// doi.org/10.3390/molecules26082105
SponsorshipARAID: MEC (CTQ2013-44367-C2-2-P, BFU2016-75633-P and PID2019-105451GBI00 to RH-G, CTQ2017-85658-R and CTQ2014-56370-R to AO); Gobierno de Aragón (E35_R20 and LMP58_18); FEDER (2014-2020) funds for ‘Building Europe from Aragón’; Juan de la Cierva fellowship IJCI-2017-32874
Protein O-fucosyltransferase 1 (PoFUT1) is a GT-B fold enzyme that fucosylates proteins containing EGF-like repeats. GT-B glycosyltransferases have shown a remarkable grade of plasticity adopting closed and open conformations as a way of tuning their catalytic cycle, a feature that has not been observed for PoFUT1. Here, we analyzed Caenorhabditis elegans PoFUT1 (CePoFUT1) conformational behavior in solution by atomic force microscopy (AFM) and single-molecule fluorescence resonance energy transfer (SMF-FRET). Our results show that this enzyme is very flexible and adopts mainly compact conformations and to a lesser extend a highly dynamic population that oscillates between compact and highly extended conformations. Overall, our experiments illustrate the inherent complexity of CePoFUT1 dynamics, which might play a role during its catalytic cycle.